1.Isolation and Identification of Staphylococcus xylosus in Nude Mice with Squamous Skin Scurfs
Zhihao KONG ; Xiaofeng WEI ; Lingzhi YU ; Liping FENG ; Qi ZHU ; Guojun SHI ; Chen WANG
Laboratory Animal and Comparative Medicine 2025;45(3):368-375
Objective To isolate pathogenic bacteria from the skin of a nude mouse exhibiting squamous skin scurfs, and perform bacterial identification, traceability analysis, and pathogenicity studies to provide a new approach for the diagnosis of pathogens in nude mice with squamous skin scurfs. MethodsSkin swab samples were collected from a nude mouse exhibiting squamous skin scurfs for nucleic acid testing, bacterial isolation and culture, biochemical identification, 16S rDNA gene amplification and sequencing, and whole genome sequencing to construct a phylogenetic tree. Fifteen BALB/c nude mice were randomized into a saline-treated control group, a high-concentration group treated with 1.8×10⁸ CFU/mL of the isolated bacterial suspension, and a low-concentration group treated with 1.8×10⁷ CFU/mL of the isolated bacterial suspension. Pathogenicity was assessed by animal infection experiments and observation of histopathological changes in skin tissue using HE staining. Results The nucleic acid test for Corynebacterium bovis was negative, excluding infection by this organism. The pathogen isolated on mannitol salt agar and blood agar, combined with Gram staining, suggested a Gram-positive Staphylococcus species. The isolated strain was identified by 16S rDNA sequencing and a fully automated microbial identification system as Staphylococcus xylosus. Phylogenetic tree analysis based on whole genome sequencing showed that the strain was most closely related to an isolate from leafy vegetables in South Korea (GenBank GCA_00207825.1). In the high-concentration group, squamous skin scurfs appeared on the head, neck, and back of nude mice on the 17th day post-infection, while in the low concentration group, similar symptoms appeared on the 20th day post-infection and gradually spread to other areas. The scaling symptoms were transient, lasting for 7 days in the high-concentration group and 3 days in the low-concentration group, after which the skin returned to normal. The infection rate was 33.33% in both the high- and low-concentration groups. No significant pathological changes were observed in the skin tissues of infected mice compared to the control group, indicating marked individual differences in the pathogenicity of the strain in nude mice. Conclusion A strain of Staphylococcus xylosus was isolated from the skin of a nude mouse exhibiting squamous skin scurfs. The strain is an opportunistic pathogen that causes transient squamous skin scurfs without significant histopathological changes, and there are individual differences in the sensitivity of nude mice to this strain. These findings can provide valuable data for pathogen identification in immunodeficient or gene knockout mice.
2.Isolation and Identification of Staphylococcus xylosus in Nude Mice with Squamous Skin Scurfs
Zhihao KONG ; Xiaofeng WEI ; Lingzhi YU ; Liping FENG ; Qi ZHU ; Guojun SHI ; Chen WANG
Laboratory Animal and Comparative Medicine 2025;45(3):368-375
Objective To isolate pathogenic bacteria from the skin of a nude mouse exhibiting squamous skin scurfs, and perform bacterial identification, traceability analysis, and pathogenicity studies to provide a new approach for the diagnosis of pathogens in nude mice with squamous skin scurfs. MethodsSkin swab samples were collected from a nude mouse exhibiting squamous skin scurfs for nucleic acid testing, bacterial isolation and culture, biochemical identification, 16S rDNA gene amplification and sequencing, and whole genome sequencing to construct a phylogenetic tree. Fifteen BALB/c nude mice were randomized into a saline-treated control group, a high-concentration group treated with 1.8×10⁸ CFU/mL of the isolated bacterial suspension, and a low-concentration group treated with 1.8×10⁷ CFU/mL of the isolated bacterial suspension. Pathogenicity was assessed by animal infection experiments and observation of histopathological changes in skin tissue using HE staining. Results The nucleic acid test for Corynebacterium bovis was negative, excluding infection by this organism. The pathogen isolated on mannitol salt agar and blood agar, combined with Gram staining, suggested a Gram-positive Staphylococcus species. The isolated strain was identified by 16S rDNA sequencing and a fully automated microbial identification system as Staphylococcus xylosus. Phylogenetic tree analysis based on whole genome sequencing showed that the strain was most closely related to an isolate from leafy vegetables in South Korea (GenBank GCA_00207825.1). In the high-concentration group, squamous skin scurfs appeared on the head, neck, and back of nude mice on the 17th day post-infection, while in the low concentration group, similar symptoms appeared on the 20th day post-infection and gradually spread to other areas. The scaling symptoms were transient, lasting for 7 days in the high-concentration group and 3 days in the low-concentration group, after which the skin returned to normal. The infection rate was 33.33% in both the high- and low-concentration groups. No significant pathological changes were observed in the skin tissues of infected mice compared to the control group, indicating marked individual differences in the pathogenicity of the strain in nude mice. Conclusion A strain of Staphylococcus xylosus was isolated from the skin of a nude mouse exhibiting squamous skin scurfs. The strain is an opportunistic pathogen that causes transient squamous skin scurfs without significant histopathological changes, and there are individual differences in the sensitivity of nude mice to this strain. These findings can provide valuable data for pathogen identification in immunodeficient or gene knockout mice.
3.Cerebrospinal fluid metagenomic next-generation sequencing for the diagnosis of intracranial aspergillus flavus infection in immunocompetent patients: A case report
Xianzhe KONG ; Huan WEI ; Liping ZHAN
Journal of Apoplexy and Nervous Diseases 2025;42(7):656-658
To report a case of an immunocompetent young adult male patient diagnosed with intracranial Aspergillus flavus infection, and to investigate the clinical features of this disease and related experience in diagnosis and treatment.A retrospective analysis was performed for the clinical data of a patient who had the initial presentation of high fever and headache and then progressed to meningoencephalitis, and the results of cerebrospinal fluid (CSF) metagenomic next-generation sequencing (mNGS) and treatment outcomes were summarized.The patient had an acute onset, with no response to empirical anti-infective therapy in the incipient stage, and then he gradually developed disturbance of consciousness and meningeal irritation sign. CSF analysis showed inflammatory changes, while conventional pathogen tests yielded negative results, and mNGS detected 27 specific sequences of Aspergillus flavus. The symptoms of the patient was significantly improved after antifungal therapy with voriconazole, with no recurrence after follow-up for 3 months.For unexplained central nervous system infections, especially those with negative results from conventional tests, mNGS can improve the detection rate of rare pathogens(e.g.,Aspergillus flavus). Early diagnosis and targeted antifungal therapy are crucial for improving prognosis. This case highlights that invasive fungal infections should be considered even in immunocompetent individuals.
Aspergillus flavus
4.Factors affecting the social alienation among the elderly patients with type 2 diabetes mellitus
ZHOU Mengjiao ; LI Meilin ; KONG Liping
Journal of Preventive Medicine 2024;36(4):292-295
Objective:
To investigate the influencing factors for social alienation among the elderly patients with type 2 diabetes mellitus (T2DM), so as to provide insights into formulating targeted intervention measures.
Methods:
T2DM patients aged 60 years and older were selected from Hangzhou First People's Hospital from January to October 2023. Their demographics, diabetes complications were collected using questionnaire surveys, social support level was evaluated using the Social Support Scale, and the stigma was evaluated using the Chinese version of the Type 2 Diabetes Stigma Assessment Scale, and the social alienation was assessed using General Alienation Scale. Factors affecting the social alienation were identified among the elderly patients with T2DM using a multiple linear regression model.
Results:
A total of 316 elderly patients with T2DM were investigated, including 171 males (54.11%) and 145 females (45.89%), and had a mean age of (69.75±8.12) years. The level of social support was mainly medium, with 162 cases accounted for 51.27%. The average stigma scores were (61.87±12.50) points, and average social alienation scores were (42.09±6.33) points. Multiple linear regression analysis identified educational level (high school/junior college, β'=-0.159; college and above, β'=-0.301), marital status (married persons, β'=-0.236), monthly household income (3 000 Yuan and more, β'=-0.175), diabetes complications (β'=0.192), social support level (medium, β'=-0.210; high, β'=-0.352) and stigma score (β'=0.283) as factors affecting the social alienation among the elderly patients with T2DM.
Conclusion
The social alienation among the elderly patients with T2DM are associated with educational level, marital status, family income, diabetes complications, social support level and stigma.
5.Correlation of health self-focused attention with knowledge, attitude and practice in health management of patients with essential hypertension
Liping CHEN ; Yuyang CAI ; Li KONG ; Fulai SHEN ; Ying JIN
Chinese Journal of General Practitioners 2024;23(1):33-40
Objective:To explore the impact of health self-focused attention on knowledge, attitude and practice of health management in patients with essential hypertension.Methods:This was a cross-sectional study. A total of 190 patients with essential hypertension were randomly selected from Shanghai Dapuqiao Community Health Service Center to participate in a questionnaire survey from October to December, 2021. The contents of the questionnaire included the general information of the patients, the degree of influence of hypertension on individuals, and the patients′ knowledge and personal cognition of hypertension, self-management behaviors, health self-focused attention, and blood pressure control. SPSS 25.0 and AMOS 24.0 were used to develop a structural equation model for analysis.Results:The recovery rate of questionnaires was 100.0% (190/190). Of the 190 patients, there were 82 males and 108 females with a mean age of (71.2±8.5) years. According to blood pressure, 190 hypertensive patients were divided into blood pressure controlled group ( n=119) and blood pressure non-controlled group ( n=71). There were significant differences in the total scores of hypertension self-management behavior and health self-focused attention between the two groups ( P<0.05). Correlation analysis showed that the degree of influence of hypertension on individuals was negatively correlated with the disease knowledge and personal cognition, and the performance of hypertension self-management behavior ( r=-0.39 and -0.28, P<0.01), while the disease knowledge and personal cognition were positively correlated with the performance of hypertension self-management behavior and health self-focused attention ( r=0.29 and 0.27, P<0.01). Hypertension self-management behavior was positively correlated with health self-focused attention and blood pressure control ( r=0.28 and 0.48, P<0.01), and health self-focused attention was positively correlated with blood pressure control ( r=0.21, P<0.01). The equation model analysis showed that health self-focused attention had a driving effect on patients′ disease knowledge and cognition and self-management behavior. Health self-focused attention significantly affected patients′ mastery of disease knowledge and formation of correct cognition (path coefficient=0.28). In addition, hypertension patients′ disease knowledge storage and internal recognition of prevention and treatment strategies determined their self-management behavior (path coefficient=0.20). On the other hand, the impact of hypertension on patients life, work and economic burden played a negative role in their acceptance of disease management knowledge, formation of correct concepts, and adoption of self-management behaviors with adjustment coefficients of -0.48 and -0.22, respectively. Conclusion:Health self-focused attention may play an initial role in the process of changing the knowledge, attitude and behavior of hypertension patients.
6.Establishment of a qPCR method to detect Staphylococcus xylosus and its application
Lingzhi YU ; Liping FENG ; Zhihao KONG ; Qi ZHU ; Xiaofeng WEI
Acta Laboratorium Animalis Scientia Sinica 2024;32(1):73-79
Objective To establish and evaluate a method for rapid and sensitive S.xylosus detection using qPCR(real-time quantitative PCR).Methods A gehM gene fragment was selected as the target for S.xylosus.A set of specific primers was synthesized and a qPCR method was established to detect S.xylosus.A S.xylosus standard strain and other non-target strains were chosen for analysis.DNA of S.xylosus was diluted 10-fold to determine its sensitivity.Clinical samples were tested,and positive products were sequenced.The result were compared with those of bacterial culture.Results S.xylosus had a specific amplification curve,whereas other non-S.xylosus species did not,indicating that the primers were specific for S.xylosus.Sensitivity was 100 fg/μL DNA.Repeatability within and between groups was less than 3%.A total of 60 clinical samples were analyzed,of which five samples had a typical S curve.qPCR products were sequenced and BLAST searched.The similarity of the gene sequences was 99.63%,indicating that the sample was positive for the S.xylosus gehM gene with a positivity rate of 8.3%.However,the positivity rate of bacterial culture was 6.7%.The positivity rate of qPCR was slightly higher than that of the culture.Conclusions The established qPCR method is rapid with high sensitivity and specificity,and can be used to detect S.xylosus.
7.Research progress in ultrasound imaging features of parathyroid diseases
Kexin MA ; Lingquan KONG ; Liping LIU ; Yuwei YANG
Chinese Journal of Endocrine Surgery 2024;18(4):586-588
Parathyroid disease is an endocrine disease caused by abnormal secretion of parathyroid hormone, which causes systemic multi-system reactions. Ultrasonography is currently the first-line method for the evaluation of parathyroid diseases and one of the important tools for preoperative localization of parathyroid lesions. In order to improve the ultrasonic diagnosis rate of parathyroid diseases, this article reviews the conventional ultrasound, contrast-enhanced ultrasound and ultrasound elastography characteristics of normal parathyroid glands and parathyroid diseases.
8.Pedigree analysis of B el subtype caused by the new allele c.175_176insGA
Hecai YANG ; Yin GUAN ; Xiaoli MA ; Yonglei LYU ; Yongkui KONG ; Chaoqun GUO ; Minglu GENG ; Liping WANG ; Tao WEN
Chinese Journal of Laboratory Medicine 2024;47(10):1206-1211
Objective:To serologically and genotypically analyze the pedigree of a case with a new allele c.175_176insGA of B el subtype and preliminarily explore the molecular mechanism of weak expression of glycosyltransferase B. Method:In the descriptive study,a 23-year-old male voluntary blood donor and his family members were selected for the study. The ABO and Le blood types of the proband and his family members was identified by the test tube method. The agglutination inhibition test was applied to detect the B and H antigens in saliva, and the Sanger sequencing and PacBio (Pacific Bioscience) third-generation haplotype sequencing were performed on the study subjects to identify genotypes. Finally, Expasy software were applied to amino acid translation of DNA sequences and prediction of protein length after gene alteration. ORF finder was applied to predict alternative start codons as well as open reading frames of mRNA, and protein expression mechanisms were analyzed.Results:The proband and her sister were B el subtype, her mother was AB el subtype, her father was normal O type, and all members of the family were Le(a+b+) phenotype. Sanger sequencing results showed that a new allele of c.175_176insGA was found in exon 4 of the proband, her mother, and her sister. Third-generation haplotype sequencing detected the haplotypes of the family members, which revealed that the proband was ABO*O.01.02/ABO*BEL.NEW (c.175_176insGA), the father was ABO*O.01.02/ABO*O.01.02, the mother was ABO*A1.02/ABO*BEL.NEW (c.175_176insGA), and the sister was ABO*O.01.02/ABO*BEL.NEW (c.175_176insGA). Analysis of the protein expression mechanism indicated that although the new allele of ABO*BEL.NEW was presumed to cause a frameshift mutation and result in a premature stop codon p.Asp59Glu*fs20 in exon 5, encoding an inactive glycosyltransferase, an alternative start codon could be utilized to initiate translation of B el subtype functional glycosyltransferase. Conclusion:Expression of the new allele of B el subtype is associated with the translation of B el subtype glycosyltransferase initiated by alternative start codons.
10.Effect of diet intervention based on IMB model in patients with diabetes combined with gout
Fangxia YIN ; Cong WANG ; Liping KONG ; Qingqing WU
Chinese Journal of Modern Nursing 2023;29(18):2467-2471
Objective:To explore the effect of diet intervention based on information-motivation-behavioral skill (IMB) model in patients with diabetes combined with gout.Methods:From January 2019 to September 2021, diabetes patients with gout admitted to the Hangzhou First People's Hospital were selected by convenience sampling. According to the random number table method, patients were divided into a control group (60 cases) and a study group (60 cases). Both groups received symptomatic treatment and routine nursing. On this basis, the control group received routine diet intervention, while the research group received diet intervention based on the IMB model. The Diet Compliance Questionnaire and the Chinese version of the World Health Organization Quality of Life Measurement Scale-Brief Version (WHOQOL-BREF) were used to evaluate the diet compliance and quality of life of two groups of patients. This study measured the blood uric acid, glycosylated hemoglobin (HbA1c), and fasting blood glucose (FPG) in two groups of patients to evaluate their health condition.Results:After intervention, the scores of knowledge, attitude and practice dimensions in the Diet Compliance Questionnaire of patients in the study group were higher than those in the control group, and the Chinese version of WHOQOL-BREF score was higher than that in the control group, with statistical differences ( P<0.05). After intervention, the blood uric acid, FPG, and HbA1c in the study group were lower than those in the control group, with statistically significant differences ( P<0.05) . Conclusions:The diet intervention based on IMB model can improve the diet compliance of diabetes patients with gout, reduce the blood glucose and uric acid of patients, and improve the quality of life of patients.


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